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Preprint Title First Total Synthesis of Hoshinoamides A

Authors Haipin Zhou, Zihan Rui, Yiming Yang, Shengtao Xu, Yutian Shaoand Long Liu

Publication Date 15 Sep 2021

Article Type Letter

Supporting Information File 1 Supporting Information Hoshinoamides A.docx; 1.2 MB

ORCID® iDs Haipin Zhou - https://orcid.org/0000-0003-2961-5829; Long Liu -https://orcid.org/0000-0003-0821-6992

1

First Total Synthesis of Hoshinoamides A

Haipin Zhoua, Zihan Ruia, Yiming Yanga, Shengtao Xua,b, Yutian Shao a,*and Long

Liuc,*

aCollege of Materials & Chemical Engineering, Chuzhou University, Chuzhou

239000, China

bState Key Laboratory of Natural Medicines and Department of Medicinal Chemistry,

China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China

cTaizhou Medical Hi-Tech Development Public Services Platform, Taizhou 225300,

China;

Email: Long Liu---15102209730@163.com

Email: Yutian Shao ---shaoyutian22@163.com

* Corresponding author

2

Abstract

Hoshinoamides A, B and C, linear lipopeptides, were isolated from the marine

cyanobacterium Caldora penicillata, with potent antiplasmodial activity against chloroquine-

sensitive Plasmodium falciparum. Herein, we describe the first total synthesis of

Hoshinoamides A. Our synthetic strategy uses the combined methods of solution and solid

phase peptide synthesis. Liquid phase synthesis is to improve the coupling yield of L-Val3 and

N-Me-D-Phe2. Connecting other amino acids efficiency and convergence by solid state

synthesis. This synthetic strategy has good purity and high yield.

Keywords

highly methylated polypeptides; Hoshinoamides; total synthesis; antimalarial;

Introduction

Malaria is an insect-borne infectious disease caused by parasites of the genus Plasmodium,

which seriously threatens human life and health[1]. Half of the world's population is at the risk

of malaria, with about 200 million new infections and killing hundreds of thousands of people

each year[2]. Current medicines for malaria include quinolone[3,4], folic acid antagonist[5,6] and

artemisinin derivatives[7]. The emergence of drug resistance makes the efficacy of these drugs

decline year by year, forcing scientists to constantly search for new antimalarial drugs[8-10].

In recent years, Iwasaki and co-workers reported three novel linear lipopeptides natural

products, Hoshinoamides A, B [11]and C[12], from a microbial metabolite of marine

cyanobacterium Caldora penicillata (Figure 1). Hoshinoamides A and B showed potent

activities against chloroquine-sensitive Plasmodium falciparum 3D7 with IC50 values of 0.52

and 1.0 μM, respectively. Hoshinoamides C inhibited the growth of the malaria parasites (IC50

3

0.96 μM ) and African sleeping sickness (IC50 2.9 μM ). Both Hoshinoamides A and B are

highly methylated polypeptides containing three N-methyl amino acids: N-Me-L-Leu7, N-Me-

D-Val5 and N-Me-D/L-Phe2. Hoshinoamides C includes two N-methyl amino acids: N-Me-D-

Phe2 and N-Me-D-IIe5. The C-terminal is Pro methyl ester while the N-terminal polypeptide

is linked to long alkyl chain amino acid Aha8/Ana8/Ama7 and p-hydroxybenzoic acid

Hba9/Hba8. Hoshinoamides have a relatively simple structure and therefore make an attractive

target for further medicinal chemistry studies. To enable these new SAR studies, we would

first need to develop efficient synthetic method to provide sufficient material. Hoshinoamides

A shows better antimalarial activity and less cytotoxicity compared to Hoshinoamides B.

Herein, we report the initial progress on the total synthesis of Hoshinomaides A.

The key challenges for the total synthesis of Hoshinoamides A are the coupling of highly

methylated amino acids and the purification of hydrophobic peptides.

4

Figure 1. a) Structures of Hoshinoamides A and B. b) Structures of Hoshinoamides C

HO

HN

O

N

OO

HN

H

N

OHN

O

NH2O

NH

O

N

O

N

OO

OMe

N

OO

HN

H

N

OHN

O

NH2O

O NN

OO

OMe

NH

OHO

O

N

Hba-Aha-N-Me-Leu-Ile-N-Me-D-Val-Gln-Val-N-Me-D-Phe-Pro-OMe

Hba-Ana-N-Me-Leu-Ile-N-Me-D-Val-Gln-Pro-N-Me-Phe-Pro-OMe

Hba9

Ana8

N-Me-L-Leu7

L-Ile6

N-Me-D-Val5

L-Gln4

L-Val3

N-Me-D-Phe2

L-Pro-OMe1

L-Pro-OMe1

N-Me-L-Phe2

L-Pro3

L-Gln4L-Ile6

Hba9

Aha8

N-Me-L-Leu7 N-Me-D-Val5

Hoshinoamide A

Hoshinoamide B

a)

b)

NH

OHO

HN

O

N

OHN

O

H

NH2O

NH

O

O

N

N

O

OMe

O

L-Pro-OMe1

N-Me-D-Phe2

L-Val3

L-Gln4

N-Me-D-Ile5

L-Leu6

Ama7Hba8

Hoshinoamide C

Hba-Ama-Leu-N-Me-D-Ile-Gln-Val-N-Me-D-Phe-Pro-OMe

5

Results and Discussion

As shown in Scheme 1, we initially tested Fmoc Solid-phase peptide synthesis (SPPS) to

get 2-chlorotrityl resin-bound Pro1-(N-Me)Phe2 dipeptide 2 under the condition of HCTU and

DIPEA. Unfortunately, the N-Me coupling proceeded in low yield (< 10%).

Scheme 1. Synthesis of resin-bound tripeptide 3 by SPPS

In order to improve the coupling yield of hindered peptide, we tried to condensation of Val3

with dipeptide in solution phase. Pro-Bn 5 was first coupled with Fmoc-N-Me-D-Phe-OH by

the treatment of HATU and DIPEA, giving the dipeptide 6 in 83% yield (Table 1). We

envisioned a sequential deprotection of Fmoc of dipeptide 6 and then coupling with Fmoc-

Val-OH will deliver tripeptide 7. With this in mind, we next screened a series of coupling

reagents. As shown in Table 1, the coupling reagents have a significant effect on the

efficiencies of the reactions. While most of coupling reagents could give the tripeptide 7, the

combination of HATU/DIPEA shown the best result, delivering 7 in 78% isolated yield

(Table 1, entry 2). Only trace product could be detected while EEDQ was used as the

coupling reagent. The purity of tripeptide 7 was determined by HPLC and no racemization

was observed, ensuring the smooth progress of the total synthesis of Hoshinoamides A

6

Table 1: Hindered Peptide coupling: Conditions and Yields

.

Entry Coupling reagenta Yieldb (%) racemizationc

1 HCTU, DIPEA 36 NO

2 HCTU, DIPEA 78d NO

3 HATU, DIPEA 75 NO

4 HATU, HOAT, DIPEA trace NO

5 EEDQ 41 NO

6 DIC, HOAT 24 NO

a0.1 mmol/L of dipeptide 6 and Fmoc-Val in DMF, 1.5 equiv coupling reagent, rt, 3 h. bYield

were determined by H NMR data analysis. cRacemization were determined by HPLC.

dIsolated yield.

With the Tripeptide 7 in hand, we went on to construct the peptide scaffold (Scheme 2).

Deprotection of the Bn groups by Pd(OH)2-catalyzed hydrogenolysis gave the tripeptide 8.

Treatment of 2-chlorotrityl (CTC) resin with 8 (4 equiv.) with DIPEA successfully produced

the resin-bound tripeptide 3 in good yield. It should be noted unreacted 8 can be largely

recovered by a quick silica gel chromatography. The N terminus of 3 was then sequentially

extended with properly protected L-Gln4, N-Me-D-Val5, L-Ile6, N-Me-L-Leu7, Ana8, and Hba9

units to give 9-mer peptide 9 using the standard SPPS procedure. It is worth noting that, in

order to increase the yield, when coupling L-Ile6 and Ana8, the reaction time increases, and

the reaction times increase. The peptide chain was then cleaved from resin by the treatment of

0.5% TFA in DCM and removal of the Trt group with aqueous solution of TFA gave 9-mer

peptide 10 in good yield after HPLC purification. The carboxylic acid of 9-mer peptide 10

was converted to ester with MeI and K2CO3 in DMF, delivering the final natural product

7

Hoshinoamides A in 2% yield (10 mg). The spectroscopic data of synthetic Hoshinoamides A

were in excellent agreement with the data previously reported for the natural product.

Scheme 2. Synthesis of Hoshinoamides A

8

Conclusion

In summary, we completed the first total syntheses of Hosh-inoamides A. Through the

combination of the solution and solid phase peptide synthesis, Hoshinoamides A was

synthesized in high efficency. After systematic screening of the coupling reagents in solution

phase, the key intermediate tripeptide 7 was obtained with high yield. The solid phase

synthesis improves the entire efficiency of the synthetic route. This strategy could be applied

to stereoselectively synthesize Hoshinoamides A and other highly methylated polypeptides

analogues, which was helpful to further study its biological activity against malaria. Structure-

and-activity and functional studies with the fluorescent-labelled analogs is currently under

investigation in our lab.

Experimental

General Experimental Procedures. 1H-NMR spectra were obtained using a Bruker

AVANCE AV 400 at frequencies of 400 MHz respectively in CDCl3, CD3OD or D2O.

Chemical shifts are reported in parts per million (ppm) and coupling constants in Hertz (Hz).

The residual solvent peaks were used as internal standards. 1H-NMR data is reported as

follows: chemical shift values (ppm), multiplicity (s = singlet, d = doublet, t = triplet, q =

quartet, m = multiplet), coupling constant and relative integral. 13C-NMR spectra were

obtained using a Bruker AVANCE AV 400 at 100 MHz in CDCl3, CD3OD or D2O. 13C-NMR

data is reported as chemical shift values (ppm).

LC-MS was performed on a Thermo Scientific MSQ instrument with the spectrometer

operating in positive mode. Separations on the LC-MS system were performed on two

methods using a thermo accucore C18 (2.6 µm, 100 x 2.1 mm) column. Method A: Linear

gradient of 10-90% CH3CN/H2O and 0.1% TFA over 40 min was applied at a flow rate of 1.0

9

mL/min and detection at 220 nm. Method B: Linear gradient of 10-90-90-10% CH3CN/H2O

and 0.1% TFA over 10 min (10-90 vol% MeCN over 6 min, 90-90 vol% over 3 min, 90-10

vol% MeCN over 1min) was applied at a flow rate of 1.0 mL/min and detection at 220 nm.

Preparative reverse-phase HPLC was performed using Thermo Scientific Ultimate 3000

equipped with a Thermo Hypersil Gold (5 µm, 150 x 21.2 mm) column using the following

buffer systems: A: 0.1% TFA in water. B: 0.1% TFA in MeCN using a 10-90-90-10 vol%

MeCN gradient (10-90 vol% MeCN over 30 min, 90-90 vol% over 10 min, 90-10 vol%

MeCN over 10 min) at a flow rate of 8 mL/min.

Standard SPPS (solid phase peptide synthesis) method:

1) General procedure for coupling on resin: The loaded resin was shaken for 2 h at room

temperature with a solution of the desired Fmoc-AA-OH (4 equiv), HATU/Coupling reagent

(4 equiv) and DIPEA (8 equiv) in DMF (20 mL). The coupling mixture was filtered and the

resin was washed with CH2Cl2 (10 mL x 5) and CH3OH (10 mL x 5).

2) General procedure for deprotection of Fmoc: The loaded resin was treated with a

solution of 20 vol% piperidine in DMF (20 mL) for 30 min and then filtered. The resin was

washed with CH2Cl2 (20 mL x 5) and CH3OH (20 mL x 5).

3) General procedure for cleavage the peptide from the resin: 0.5% TFA in DCM (20 mL)

were added on the resin and the mixture was shaken for 2h before filtered. The resin was

washed with CH2Cl2 (20 mL x 5) and CH3OH (20 mL x 5).

Fmoc-Pro-OH (674 mg, 2 mmol) was then dissolved in a mixture of DCM (10 mL) and

DMF (10 mL). DIPEA (1.7 mL, 10 mmol), 2-CTC resin (1 g) were added to this mixture and

the reaction was stirred at room temperature was for 2h. The resin was filtered and washed

with MeOH (3 x 20 mL), DCM (3 x 20 mL). The unreacted resin was capped with MeOH in a

mixture of MeOH:DIPEA:DCM (1:2:7, 10 mL) for 3h. The resin-bound peptide was added to

a mixture of 20% piperidine in DMF (20 mL), and the mixture was shaken to for 30 minutes.

Then the mixture was filtered, the resin was washed with MeOH (3 x 20 mL) and DCM (3 x

10

20 mL). Fmoc-N-Me-D-Phe-OH (1000 mg, 2.5 mmol), HCTU (1.0 g, 2.5 mmol) and DIPEA

(871 µL, 5.0 mmol) in DMF were added on the resin and the reactor was shaken for 1h at

room temperature. Then the mixture was filtered, the resin was washed with MeOH (3 x 20

mL) and DCM (3 x 20 mL) to afford the resin-bound dipeptide. The resin-bound dipeptide

was added to a mixture of 20% piperidine in DMF (20 mL), and the mixture was shaken to for

30 minutes. Then the mixture was filtered, the resin was washed with MeOH (3 x 20 mL) and

DCM (3 x 20 mL). Fmoc-Val-OH (848 mg, 2.5 mmol), HATU (1.0 g, 2.5 mmol) and DIPEA

(871 µL, 5.0 mmol) in DMF were added on the resin and the reactor was shaken for 1h at

room temperature. The resulting tripeptide was analysed on a Thermo Scientific MSQ

instrument, and few product was observed.

Fmoc-N-Me-D-Phe-Val-Pro-Bn (6). Pro-OBn.HCl (2.41 g, 10 mmol), Fmoc-N-Me-D-

Phe-OH (4.01 g, 10 mmol) and DIPEA (5.2 mL, 30 mmol) was dissolved in 50 mL anhydrous

DCM. HATU (5.7 g, 15 mmol) was added to the solution and the mixture was stirred at room

temperature for 6 h. The reaction mixture was then washed by 1.0 M HCl (20 mL), aqueous

NaHCO3 (20 mL) and brine (20 mL). The organic phase was dried with anhydrous Na2SO4

and concentrated in vacuo. The crude residue was purified by flash column chromatography

(n-hexanes: EA=2:1) to afford dipeptide 6 (4.9 g, 83%).1H NMR (400 MHz, Methanol-d4) δ

7.39 – 7.27 (m, 19H), 5.20 – 5.12 (m, 4H), 4.85 (s, 6H), 4.44 (ddd, J = 13.1, 9.5, 4.8 Hz, 4H),

3.50 (ddd, J = 10.0, 7.4, 5.4 Hz, 2H), 3.31 – 3.23 (m, 4H), 3.07 (dd, J = 12.8, 10.3 Hz, 2H),

2.57 (s, 6H), 2.46 (dt, J = 9.9, 7.0 Hz, 2H), 2.03 – 1.94 (m, 2H), 1.89– 1.71 (m, 4H), 1.48

(dddd, J = 12.5, 7.1, 5.4, 1.6 Hz, 2H). 13C NMR (101 MHz, MeOD) δ 171.21, 165.90, 135.71,

133.51, 129.37, 128.70, 128.24, 128.07, 127.93, 127.74, 66.74, 60.75, 59.32, 48.27, 48.06,

47.84, 47.63, 47.41, 47.21, 46.99, 46.88, 36.55, 30.88, 28.45, 23.94. HRMS: (+ESI) Calc. for

C22H26N2O3: 588.2671[M+H]+, Found: 588.2673 [M+H]+.

11

Fmoc-Val-N-Me-D-Phe-Val-Pro-Bn (7). Dipeptide 6 (118 mg, 0.20 mmol), Fmoc-Val-

OH (71 mg, 0.20 mmol) was dissolved in 10 mL anhydrous DMF. Coupling reagents was

added to the solution and the mixture was stirred at room temperature for 3 h. This mixture

poured onto water (10 mL) and extracted with CH2Cl2 (3 x 10 mL). Then washed by 1.0 M

HCl (10 mL), aqueous NaHCO3 (10 mL) and brine (10 mL). The organic phase was dried

with anhydrous Na2SO4 and concentrated in vacuo. The crude residue was purified by flash

column chromatography (n-hexanes: EA=2:1) to afford tripeptide 7. 1H NMR (400 MHz,

Chloroform-d) δ 7.75 (d, J = 7.7 Hz, 2H), 7.57 (t, J = 7.2 Hz, 2H), 7.40 – 7.17 (m, 14H), 5.72

(dd, J = 8.9, 6.5 Hz, 1H), 5.46 (d, J = 9.5 Hz, 1H), 5.23 – 5.20 (m, 1H), 5.06 (d, J = 12.2 Hz,

1H), 4.50 – 4.18 (m, 5H), 3.48 – 3.43 (m, 1H), 3.28 (dt, J = 11.3, 5.8 Hz, 2H), 3.10 (s, 1H),

2.95 – 2.86 (m, 3H), 2.21 – 2.14 (m, 2H), 1.78 – 1.62 (m, 5H), 1.28 (s, 2H), 0.76 (dd, J =

66.2, 6.8 Hz, 3H), 0.47 (dd, J = 46.7, 6.7 Hz, 3H). 13C NMR (101 MHz, CDCl3) δ 171.70,

171.60, 168.26, 156.39, 143.88, 141.30, 137.01, 129.54, 128.88, 128.69, 128.60, 128.50,

128.39, 128.31, 128.20, 127.72, 127.08, 126.65, 125.17, 125.08, 119.99, 77.40, 77.09, 76.77,

67.04, 66.84, 59.43, 55.87, 55.59, 47.17, 46.92, 35.00, 30.72, 30.47, 28.78, 25.25, 19.82,

16.33. HRMS: (+ESI) Calc. for C42H45N3O6: 688.3381 [M+H]+, Found: 688.3384[M+H]+.

Comparison of the effects of different coupling reagents on the reaction yield (Table 1).

Fmoc-Val-N-Me-D-Phe-Val-Pro-OH (8). Tripeptide 7 (2.3 g, 3.3 mmol) was was

dissolved in 30 mL of MeOH/HCOOH(v/v=9:1) and hydrogenized with Pd(OH)2 (500 mg,

10% on carbon) under H2 for 10 hours to remove the Bn groups. The reaction mixture was

filtered through a pad of celite and the filtrate was concentrated in vacuo to give brown oil

which was purified by flash chromatography (n-hexanes:EA = 2:1), affording tripeptide 8

(1.87 g, 95%) as a white foam. 1H NMR (400 MHz, Methanol-d4) δ 7.59 (d, J = 8 Hz, 2H),

7.45 (d, J = 8 Hz, 2H), 7.21 – 6.96 (m, 10H), 5.52 (ddd, J = 43.6, 9.9, 5.6 Hz, 1H), 4.81 (s,

2H), 4.20 – 3.98 (m, 4H), 3.29 – 3.10 (m, 2H), 3.04– 2.91 (m, 4H), 2.80 – 2.67 (m, 2H), 2.62

12

(s, 3H), 2.01 – 1.83 (m, 2H), 1.53 (ddq, J = 50.8, 19.7, 6.9 Hz, 4H), 1.13 – 1.09 (m, 2H), 0.52

– 0.45 (m, 4H). 13C NMR (101 MHz, MeOD) δ 175.49, 175.06, 174.95, 173.81, 173.14,

171.16, 170.16, 167.42, 164.85, 158.64, 158.58, 158.53, 145.38, 145.35, 145.20, 145.13,

145.09, 142.88, 142.55, 138.50, 138.41, 131.03, 130.75, 130.66, 130.27, 129.77, 129.64,

129.57, 129.42, 128.86, 128.83, 128.55, 128.28, 128.21, 127.77, 127.66, 126.35, 126.29,

126.21, 125.76, 121.02, 70.63, 68.11, 68.04, 66.85, 60.78, 60.68, 59.15, 58.19, 57.76, 57.67,

57.41, 57.06, 56.41, 56.08, 49.75, 49.54, 49.32, 49.11, 48.90, 48.68, 48.47, 48.41, 48.25,

47.74, 38.96, 37.58, 37.01, 36.60, 35.79, 35.61, 33.11, 32.20, 31.89, 31.73, 31.53, 31.40,

30.88, 30.83, 30.66, 30.53, 30.39, 29.98, 29.64, 28.18, 26.97, 26.10, 23.80, 23.34, 19.88,

19.48, 18.17, 18.07, 17.64, 14.58. HRMS: (+ESI) Calc. for C35H39N3O6: 598.2912 [M+H]+,

Found: 598.2915[M+H]+.

Compound tripeptide 8 (1.20 g, 2 mmol) was then dissolved in a mixture of DCM (10

mL) and DMF (10 mL). DIPEA (1.7 mL, 10 mmol), 2-CTC resin (1 g) were added to this

mixture and the reaction was stirred at room temperature was for 3h. The resin was filtered

and washed with MeOH (3 x 20 mL), DCM (3 x 20 mL). The unreacted resin was capped

with MeOH in a mixture of MeOH:DIPEA:DCM (1:2:7, 10 mL) for 5 h. Fmoc protecting

group was removed following the general procedure and the remain amino acids were

successively coupled using the standard SPPS method. 0.5% TFA in DCM (20 mL) were

added on the resin and the mixture was shaken for 2h to cleavage the peptide from the resin.

The mixture was filtered and the filtrate was concentrated in vacuo to give a white foam. The

peptide was re-dissolved in a mixture of TFA:Et3SiH:H2O (10 mL, 50/50/50 v/v/v). The

reaction mixture was stirred for 3 h, and then concentrated in vacuo. The crude peptide was

precipitated using cold Et2O and centrifuged at 7000 rpm to give a white solid. This solid was

further purified by RP-HPLC using protocols described in the general method. Fractions were

collected, concentrated and lyophilized to give nanopeptide 10 as a white solid. Nanopetide

13

10 was dissolved in dry DMF (5 mL). K2CO3 (3.1 mg, 0.022 mmol) and MeI (3.13 mg, 0.022

mmol) was added to this solution. The reaction mixture was stirred for 3 h. This mixture

poured onto water (5 mL) and extracted with CH2Cl2 (3 x 5 mL). Then washed by 1.0 M HCl

(10 mL), aqueous NaHCO3 (10 mL) and brine (10 mL). The organic phase was dried with

anhydrous Na2SO4 and concentrated in vacuoto give brown oil . This oil was further purified

by RP-HPLC using protocols described in the general method. Fractions were collected,

concentrated and lyophilized to give Hoshinoamides A as a white solid.(10 mg, 2% yield).

The 1H NMR and 13C NMR spectra of synthetic product were fully consistent with the data of

isolated samples reported in the literature.4 See table S2 and table S3 for details. 1H NMR

(400 MHz, Methanol-d4) δ 7.30 – 7.11 (m, 6H), 7.02 – 6.94 (m, 2H), 6.72 – 6.65 (m, 2H),

5.73 (dd, J = 9.2, 6.4 Hz, 1H), 4.82 – 4.73 (m, 1H), 4.64 – 4.53 (m, 2H), 4.43 – 4.25 (m, 2H),

3.69 (s, 2H), 3.50 (dt, J = 11.2, 6.0 Hz, 1H), 3.41 – 3.31 (m, 1H), 3.20 – 3.06 (m, 9H), 2.99 –

2.92 (m, 3H), 2.91 – 2.86 (m, 1H), 2.52 (t, J = 7.6 Hz, 2H), 2.47 – 2.38 (m, 2H), 2.29 – 2.22

(m, 3H), 2.18 (q, J = 7.7 Hz, 3H), 2.05 – 1.99 (m, 1H), 1.99 – 1.71 (m, 9H), 1.68 – 1.46 (m,

6H), 1.46 – 1.34 (m, 4H), 1.11 – 0.97 (m, 3H), 0.97 – 0.81 (m, 16H), 0.68 – 0.55 (m, 6H). 13C

NMR (101 MHz, MeOD) δ 177.43, 176.52, 176.00, 174.55, 173.86, 173.15, 173.09, 172.87,

171.89, 170.47, 156.60, 138.40, 133.75, 130.69, 130.40, 129.45, 127.70, 116.18, 64.21, 60.84,

57.23, 55.69, 55.64, 55.52, 54.32, 52.69, 49.68, 49.46, 49.25, 49.25, 49.04, 48.83, 48.81,

48.61, 48.40, 40.23, 38.43, 37.82, 36.62, 35.76, 35.51, 34.59, 32.62, 31.80, 31.59, 31.56,

30.22, 29.98, 29.20, 27.72, 26.21, 26.17, 26.06, 25.95, 25.90, 25.11, 23.80, 22.17, 20.39,

19.90, 19.49, 18.13, 16.50, 11.60. HRMS: (+ESI) Calc. for C61H95N9O12: 1146.7173 [M+H]+,

Found: 1146.7173 [M+H]+

Supporting Information

Supporting Information File 1:

14

File Name: Supporting Information Hoshinoamides A

File Format: Word

Title: Supporting Information ---First Total Synthesis of Hoshinoamides A

Acknowledgements

This research was supported by the Natural Science key Project of Anhui Education

Department (Grant No. KJ2019A0635); Provincial University Students’ Innovation and

Entrepreneurship Program (Grant No. 2019CXXL077); Hubei Province Key Laboratory of

Occupational Hazard Identification and Control ， Wuhan University of Science and

Technology (Grant No. OHIC2020Y06 )

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